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( Huang Yuan Kai ),( Yao Sheng Chen ),( De Lin Mo ),( Pei Qing Cong ),( Zu Yong He ) 한국미생물 · 생명공학회 2012 Journal of microbiology and biotechnology Vol.22 No.3
Xylanase has been used extensively in the industrial and agricultural fields. However, the low-yield production of xylanase from native species cannot meet the increasing demand of the market. Therefore, improving the heterologous expression of xylanase through basic gene optimization may help to overcome the shortage. In this study, we synthesized a high-GC-content native sequence of the thermostable xylanase gene xynB from Streptomyces olivaceoviridis A1 and, also designed a slightly AT-biased sequence with codons completely optimized to be favorable to Pichia pastoris. The comparison of the sequences` expression efficiencies in P. pastoris X33 was determined through the detection of single-copy-number integrants, which were quantified using qPCR. Surprisingly, the high GC content did not appear to be detrimental to the heterologous expression of xynB in yeast, whereas the optimized sequence, with its extremely skewed codon usage, exhibited more abundant accumulation of synthesized recombinant proteins in the yeast cell, but an approximately 30% reduction of the secretion level, deduced from the enzymatic activity assay. In this study, we developed a more accurate method for comparing the expression levels of individual yeast transformants. Moreover, our results provide a practical example for further investigation of what constitutes a rational design strategy for a heterologously expressed and secreted protein.
Yuan-Hwa Chou,Wen-Sheng Huang,Ju-Wei Hsu,Shin-Min Lee,An-Shoei Yang,Kai-Chun Yang 대한정신약물학회 2010 CLINICAL PSYCHOPHARMACOLOGY AND NEUROSCIENCE Vol.8 No.2
Objective: Previous single photon emission computed tomography (SPECT) studies have demonstrated that co-injection of radiotracers 99mTc-TRODAT and 123IBZM can assess both presynaptic, dopamine transporters (DATs), and postsynaptic, dopamine D2/D3 receptors, markers of dopaminergic system simultaneously. The aim of this study was to determine both DATs and dopamine D2/D3 receptors in drug naïve schizophrenic patients. Methods: Severn drug naïve schizophrenic patients and eleven age-matched healthy controls were recruited. Each subject received one SPECT measurement after intravenous co-administration of 99mTc-TRODAT and 123IBZM and one brain MRI scan. A ratio equilibrium model was applied to calculate the specific uptake ratio (SUR) which corresponds to the ratio of Bmax (receptor density) and KD (affinity). Results: The results showed that the SUR of DATs binding was not significantly different between healthy controls and drug naïve schizophrenic patients (2.92±0.62 vs. 2.65±0.27, p=0.052) and, however, the SUR of dopamine D2/D3 receptors binding was significantly lower in drug naïve schizophrenic patients than those in healthy controls (2.79±0.54 vs. 3.90±0.58, p=0.001). Additionally, there was a significant correlation between left and right binding for both DATs (r=0.80, p=0.003) and dopamine D2/D3 receptors (r=0.96, p=0.000) in healthy controls but not seen in drug naïve schizophrenic patients (r=0.54, p=0.20 for DATs, r=0.70, p=0.08 for dopamine D2/D3 receptors). Conclusion: This study provides evidences to support the idea that abnormalities of dopaminergic system in schizophrenia and further elucidates the disruption of inter-hemispheric interaction of postsynaptic and presynaptic markers.
Fupeng Li,Kai Huang,Jinbing Wang,Kai Yuan,Yiqi Yang,Yihao Liu,Xianhao Zhou,Keyu Kong,Tao Yang,Jian He,Chunjie Liu,Haiyong Ao,Fengxiang Liu,Qian Liu,Tingting Tang,Shengbing Yang 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00
Background Although biomedical implants have been widely used in orthopedic treatments, two major clinical challenges remain to be solved, one is the bacterial infection resulting in biofilm formation, and the other is aseptic loosening during implantation due to over-activated osteoclastogenesis. These factors can cause many clinical issues and even lead to implant failure. Thus, it is necessary to endow implants with antibiofilm and aseptic looseningprevention properties, to facilitate the integration between implants and bone tissues for successful implantation. To achieve this goal, this study aimed to develop a biocompatible titanium alloy with antibiofilm and anti-aseptic loosening dual function by utilizing gallium (Ga) as a component. Methods A series of Ti-Ga alloys were prepared. We examined the Ga content, Ga distribution, hardness, tensile strength, biocompatibility, and anti-biofilm performance in vitro and in vivo. We also explored how Ga3+ ions inhibited the biofilm formation of Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) and osteoclast differentiation. Results The alloy exhibited outstanding antibiofilm properties against both S. aureus and E. coli in vitro and decent antibiofilm performance against S. aureus in vivo. The proteomics results demonstrated that Ga3+ ions could disturb the bacterial Fe metabolism of both S. aureus and E. coli, inhibiting bacterial biofilm formation. In addition, Ti-Ga alloys could inhibit receptor activator of nuclear factor-κB ligand (RANKL)-dependent osteoclast differentiation and function by targeting iron metabolism, then suppressing the activation of the NF-κB signaling pathway, thus, showing their potential to prevent aseptic loosening. Conclusion This study provides an advanced Ti-Ga alloy that can be used as a promising orthopedic implant raw material for various clinical scenarios. This work also revealed that iron metabolism is the common target of Ga3+ ions to inhibit biofilm formation and osteoclast differentiation.
Youmei Xiao,Zhanxue Xu,Yuan Cheng,Rufan Huang,Yuan Xie,Hsiang‑i Tsai,Hualian Zha,Lifang Xi,Kai Wang,Xiaoli Cheng,Yanfeng Gao,Changhua Zhang,Fang Cheng,Hongbo Chen 한국생체재료학회 2023 생체재료학회지 Vol.27 No.00
Background Ferroptosis, iron-dependent cell death, is an established mechanism for cancer suppression, particularly in hepatocellular carcinoma (HCC). Sorafenib (SOR), a frontline drug for the treatment of HCC, induces ferroptosis by inhibiting the Solute Carrier family 7 member 11 (SLC7A11), with inadequate ferroptosis notably contributing to SOR resistance in tumor cells. Methods To further verify the biological targets associated with ferroptosis in HCC, an analysis of the Cancer Genome Atlas (TCGA) database was performed to find a significant co-upregulation of SLC7A11 and transferrin receptor (TFRC), Herein, cell membrane-derived transferrin nanovesicles (TF NVs) coupled with Fe3+ and encapsulated SOR (SOR@TF-Fe3+ NVs) were established to synergistically promote ferroptosis, which promoted the iron transport metabolism by TFRC/TF-Fe3+ and enhanced SOR efficacy by inhibiting the SLC7A11. Results In vivo and in vitro experiments revealed that SOR@TF-Fe3+ NVs predominantly accumulate in the liver, and specifically targeted HCC cells overexpressing TFRC. Various tests demonstrated SOR@TF-Fe3+ NVs accelerated Fe3+ absorption and transformation in HCC cells. Importantly, SOR@TF-Fe3+ NVs were more effective in promoting the accumulation of lipid peroxides (LPO), inhibiting tumor proliferation, and prolonging survival rates in HCC mouse model than SOR and TF- Fe3+ NVs alone. Conclusions The present work provides a promising therapeutic strategy for the targeted treatment of HCC.
Chu, Jia-Qi,Jing, Kai-Peng,Gao, Xiang,Li, Peng,Huang, Rui,Niu, Yan-Ru,Yan, Shou-Quan,Kong, Jun-Chao,Yu, Cai-Yuan,Shi, Ge,Fan, Yi-Ming,Lee, Young-Ha,Zhou, Yu,Quan, Juan-Hua Landes Bioscience 2017 Cell Cycle Vol.16 No.5
<P>Autophagy and apoptosis are critical for controlling Toxoplasma gondii (T. gondii) infection. T. gondii infection during pregnancy can damage the fetus and cause birth defects; however, the molecular mechanisms of this process are poorly understood. This study aims to determine the activities of autophagy and apoptosis as well as their regulatory mechanisms during T. gondii infection by using human umbilical cord mesenchymal stem cells (hUC-MSCs) as a model of congenital diseases. LC3B, a hallmark protein of autophagy was incrementally upregulated with the infection duration, whereas p62 was downregulated in T. gondii-infected hUC-MSCs. Concurrent to this result, the invasion of T. gondii into hUC-MSCs increased in a time-dependent manner. The expression levels of Bcl-2 family proteins including Bcl-2, Bcl-xL, Bim, Bax, Bid and Bak were not altered; however, Mcl-1 levels in hUC-MSCs were dramatically decreased upon T. gondii infection. In addition, at 24h post-infection, cleaved PARP and cleaved caspase-3 protein levels were elevated in hUC-MSCs. Importantly, Mcl-1 overexpression reduced the levels of autophagy- and apoptosis-related proteins in T. gondii-infected hUC-MSCs. Mcl-1 proteins were primarily expressed in the fraction containing mitochondria and strongly interacted with Beclin-1 under normal conditions; however, these interactions were remarkably attenuated by T. gondii infection. These results suggest that mitochondrial Mcl-1 is an essential signaling mediator regulating the activation of autophagy and apoptosis during T. gondii infection.</P>